Study On Culture Conditions Optimization And Content Analyzing Of Flavonoids In Cell Culture Of Liquorice

Large-scale cell culture of Liquorice is considered to be one of the most promising alternative methods to produce its secondary metabolite. The main contents of this work were shown as follows:1. inducement of Callus, establishment of different cell line and its assessment were studied. The results showed that somatic embryonic cell was the causes of different status of callus and content of flavonoids. Light promoted the callus growth and the flavonoids accumulation. The growth curve was"S"type, and the growth cycle was about 32 days.2. The component of the culture was analyzed and the method for determination of flavonoids in culture was established. Researching conclusion indicated that glycyrrhizic acid was not found in culture and the flavonoids was composed of flavonol, flavone and flavanone et al, without the functional group of ortho-substituting hydroxyl.3. The time-courses of cell growth and flavonoids accumulation in suspension cultures were studied. It was showed that the growth curve was"S"type and growth cycle was about 20 days. The biomass had a positive correlation with the production of flavonoids, but the content dropped during the whole process. Morphological experiment indicated that the morphologic change of the cell resulted in the dropping of content of flavonoids.4. The characters of physiology and biochemistry of Liquorice were investigated. In the MS medium, sucrose was ingested quickly, and reducing sugar was consumed in the end, but a little of soluble total sugar was left. Phosphate and nitrate met the needs basically. It was showed that the pH value of suspension cultures declined in the beginning, then rise, and steadied in 5.1. The content of soluble protein rise at first then dropped, and had a linearly correlation with the biomass in the exponential phase.5. Effects of several physiological factors on the growth of cell suspension culture and flavonoids accumulation were studied. The results showed that 2,4-D had an apparent influence on the improving of biomass. MS or B5 medium supplemented with initiative pH5.8-7.0, sucrose 50g/L, nitrogen 120 mmol/L, NH4+/NO3- rate10/50, phosphate 6.0 mmol/L and Ca2+ 2.5 mmol/L were fit for the growth of suspension culture cell and flavonoids accumulation. Blue light was better for the content improvement of flavonoids.